Abstract The goals for the Multiphoton Microscopy Core Facility are to make advancements on projects that were initiated during the first two phases of the COBRE program and to expand the use of the facility to investigators that require multiphoton microscopy for novel applications, but have limited experience with these techniques. COBRE investigators, in collaboration with other faculty at the Neuroscience Center of Excellence (NCE) have used our multiphoton imaging capabilities to make discoveries in hair cell synapses and plasticity of intrinsic and synaptic properties of dendrites in the hippocampus. The pioneering multiphoton imaging equipment and its applications attracted collaborations that included scientists from outside the Center and University, including Tulane University Medical School. These diverse projects illustrate the applications that can be considered with organized collaboration and core infrastructure. While extending the availability of multiphoton imaging to more researchers in the greater New Orleans area, ultimately the core will attract sufficient new funding, which, in addition to the institutional commitment of the School of Medicine, will make the core self-sufficient by the end of Phase III funding. The three specific aims of this core are: 1) To provide the infrastructure to sustain existing grant projects that listed the core as essential equipment in their NIH applications; 2) To utilize the multiphoton facility to implement new research projects; enable collaborations between investigators; create new imaging applications in neuroscience; and help investigators obtain preliminary data for individual grant applications; 3) To develop new research strategies by continuing to develop novel capabilities including optogenetics and in vivo imaging.